Tumor necrosis factor (TNF)- plays a pro-inflammatory role in brain diseases. The biologic TNF inhibitors (TNFI), such as the TNF decoy receptor cannot be developed for brain diseases, because the TNFIs are large molecules that do not cross the blood-brain barrier (BBB). The present work continues the drug development of a re-engineered form of the human type II TNF receptor (TNFR), wherein the TNFR is produced as an IgG fusion protein. The IgG part is a genetically engineered monoclonal antibody (MAb) against the human insulin receptor (HIR). The HIRMAb part of the HIRMAb-TNFR fusion protein acts as a molecular Trojan horse to ferry the fused decoy receptor across the BBB via receptor-mediated transport on the endogenous BBB insulin receptor. The pre-SBIR feasibility stage of this research describes the engineering, expression, biochemical validation, and in vivo plasma pharmacokinetics and BBB transport in the Rhesus monkey of the HIRMAb-TNFR fusion protein. The proposed phase I SBIR research will develop a manufacturing scheme for production of the HIRMAb-TNFR fusion protein. This manufacturing will be designed to produce a therapeutic product that meets FDA specifications with regard to purity, potency, safety, and impurities, so that the manufacturing can be replicated in future GMP production of the fusion protein for clinical trials. The small scale manufacturing methodology developed in phase I will then be scaled up in phase II to the 50L bioreactor stage for manufacturing of the HIRMAb-TNFR fusion protein at production levels that can support future clinical trials. PUBLIC HEALTH RELEVANCE: Biologic tumor necrosis factor inhibitors (TNFI), such as the tumor necrosis factor (TNF) decoy receptor, cannot be developed for brain diseases, because these large molecule drugs do not cross the blood-brain barrier (BBB). The present research will re-engineer the human type II TNF receptor (TNFR) as an IgG-TNFR fusion protein, where the IgG part is a genetically engineered monoclonal antibody that crosses the BBB via transport on the endogenous insulin receptor. The IgG acts as a molecular Trojan horse to ferry across the BBB the TNFI.